Supplemental Figure 1: CIP2A and SET levels are increased in some. primary human pancreatic cancer samples. (A) CIP2A mrna levels were

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1 Supplemental Figure Legends and Figures: Supplemental Figure 1: CIP2A and SET levels are increased in some primary human pancreatic cancer samples. (A) CIP2A mrna levels were measured in 3 benign (non-cancer) pancreatic lesions and 6 ductal adenocarcinoma pancreatic samples by qrt-pcr in human patient (Pt) samples and graphed relative to the average of the 3 benign samples. (B) SET mrna levels were measured in samples from A. The average of three normals and their standard error is shown. Supplemental Figure 2: Increased expression of PP2A inhibitors trend toward decreased PP2A activity, which trends toward increased Myc expression and Myc protein stabilization in pancreatic cancer cell lines. (A) PP2A activity from Fig. 2A and total combined CIP2A and SET protein levels from Fig. 1D were used to calculate a correlation coefficient (r) using Prism GraphPad. (B) Reduced PP2A activity in pancreatic cancer cell lines trends toward increased c-myc and ps62-myc levels. PP2A activity from Fig. 2A and c- Myc and ps62-myc protein levels from Fig. 2B were used to calculate correlation coefficients (r) using Prism GraphPad. (C) c-myc and ps62-myc protein levels from Fig. 2B and combined CIP2A and SET protein levels from Fig. 1D were used to calculate correlation coefficients (r) using Prism GraphPad. (D) c-myc mrna levels are increased in some pancreatic cancer cell lines. qrt-pcr for c- Myc in the normal DT and 9 pancreatic cancer cell lines was performed and 1

2 results graphed relative to DT cells. (E) Increased c-myc protein stability in human pancreatic cancer cell lines. Myc half-life (Myc t 1/2 ) determinations were made as described in Methods for the normal DT cells and 9 pancreatic cancer cell lines. Representative immunoblots for c-myc in these experiments are shown. The calculated average c-myc half-life and standard error is shown below each immunoblot. CHX = cycloheximide. Figure Statistics: statistical analysis was carried out as described in the Materials and Methods. Error bars represent standard error. One asterisks (*) indicates a p value of while two asterisks (**) indicates a p value of less than Supplemental Figure 3: OP449 treatment decreases c-myc DNA binding and half-life in pancreatic cancer cell lines, and SET knockdown reduces OP449 sensitivity. (A) OP449 sensitivity trends with reduced SET levels. IC 50 s were determined for the indicated cell lines (see Fig. 5A) and SET protein levels from Fig. 1D were used to calculate a correlation coefficient (r) using Prism GraphPad. (B) OP449 treatment reduces c-myc target gene promoter binding. Cells were treated with OP449 or PBS as in Fig. 5B and qchip for c-myc was performed on the E2F2 and p21 promoters. (C) OP449 treatment reduces c-myc protein halflife. c-myc half-life determinations were made as described in Methods with or without OP449 treatment. Graphs showing c-myc protein remaining in these experiments (top) and western blots (bottom) are shown. Half-life measurements (Myc t 1/2 ) and standard error are indicated below each blot. CHX=cycloheximide. (D) Knockdown of SET reduces sensitivity of pancreatic cancer cells to OP449. 2

3 Stable CAPAN1 SET knockdown or control cells (immunoblot shown as inset) were plated into 96 well plates, treated with PBS or OP449 at varying doses, and population growth rates were measured using an Incucyte Zoom. Figure statistics: statistical analysis was carried out as described in the Materials and Methods. Error bars represent standard error. One asterisk (*) indicates a p value of Supplemental Figure 4: OP449 treatment reduces pancreatic cancer cell migration in vitro, proliferation in vivo, and increases tumor cell apoptosis in vivo. (A) OP449 treatment reduces pancreatic cancer cell migration. Migration assays with or without OP449 treatment were performed with Mitomycin C treated CAPAN1 and As-PC1 cells using an Incucyte Zoom, as described in Materials and Methods. The relative wound density was calculated by dividing the density inside the wound by the density outside the wound using Incucyte software. (B) A representative image from CAPAN1 cells in A is shown. (C) CAPAN1 and CFAPC1 cells were xenografted into the flank of immune compromised mice and treated with PBS or OP449 3 times per week (see Fig. 6B). After sacrifice, tumors were stained for Ki67. Representative images are shown. Figure statistics: statistical analysis was carried out as described in the Materials and Methods. One asterisks (*) indicates a p value of while two asterisks (**) indicates a p value of less than

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